Ten barley elite lines and cultivars were studied to determine the influence of genotypes, different seed cutting, media and plant hormones on the embryo-derived callus induction and plant regeneration. The results indicated that the callus induction rate of seed cutting longitudinally was higher than that of seed cutting laterally. Modified MS improved callus formation. Barley cultivars showed different response to plant hormones 2,4-D and Dicamba. The induced primary callus ultimately yielded two types of embryonic callus after subculture of 3 cycles. Regeneration media added different concentration of organic nutrients influenced plant regeneration of different barley genotypes. Albinos were observed in several cultivars after long-time subculture. Callus could be induced from all the tested cultivars, however, plant regeneration was only obtained in several genotypes. Among the ten barley cultivars 87-3175,87-0053,97-4010,97-6004 and 208813-509 were identified with good performance in callus induction and plant regeneration.